Record numbers for your first ever work but I get away for more right from your drugs out your most drugs because they are no good at least not give you something. Phase one trials by what they don't work as a last cycle and so to keep question what is the nature of the one you're out in the design process to discover that not only does a tree disease X. Y. or worse not and you know why your ideas are just right and it's you or I can. And. You live if you don't have one. And I was ever. Thank you very much for your kind hyperboles. And it's very exciting to be here today with this very friendly and yet high powered institution. So I'm going to as a title in the Kates I'm going to talk about the control of the rock specificity which as you know is a major problem in the drug discovery and quite a nightmare in the in the in their force a Big Pharma us. So from a rather oversimplified perspective there are two core problems. One has to deal when addressing side effects. One is the fact that proteins evolve from each other and hence they are the target is typically part of a family of proteins that share a common ancestor and hence this structures are uncanny really similar right in the for members of the family. They are the so-called Parra logs. So there's there's there's a great structural similarity which makes it difficult to be selective. When when many things that you don't want to keep look very much like your target. And the second major problem is that the target protein places. Roles in different similar context means it may be OK to hit a particular target or to inhibit a particular biochemical function in one type of tissue or cell but not in another and of course that may could end up causing side effects the context dependence of these target rolls. So in regards to the structural aspects which relate to the first problem. Here is an illustration of the type of problems that the community is facing when trying to develop effective drugs so for example you have on that this play out aligned the structure of two proteins two kindnesses two important signal transducers of a cell one the focal and he's and kindness. Which is a major target in cancer metastasis right. I mean that's the transducer of. Of signals and the other is essentially an receptor kinase which it's functionally completely unrelated to the former but structurally very similar in particular in the A.T.P. binding side and in fact there's a high level of amino acid conservation in the region where the competitive inhibitor is going to be targeting right. And so you can imagine that if you want to achieve specificity. You would be walking on thin ice because you want to hit the focal occasion kinase but you never want to touch the insulin receptor kinase because that would induce diabetic comma in the patient right. So that's absolutely just severely compromised. Glucose uptake of their ether of sight and so you don't really want to touch the engine receptors so how do you tell them apart if they're structurally so similar. And in fact members of the same family within the Super family. So one day I just decided to try to answer the question What is it that this thing wishes Parra logs and we can take advantage of to make more specific drugs. And it occurs to me that the. This structure is not going to be the answer but maybe some structure structural attribute which can be derived from structure provides a clue so I just decided that we probably needed to correct arise the quality of the fault. So to practice may have a perfectly aligned structure if they're close close enough homologs but the quality of the fault as the term mean by the way the site chains protect their structure may be quite different protect the structure from what from what their height from hydration of the back bone which is of course the major of the stubby lysine force of the over ninety fault. So. That takes me to really examine hydrogen bonds not so much as a P.R. wise interaction but more in regards to the environment where they are formed as you can imagine the hydrogen bond form which is mostly a little statics it's seventy eight times stronger if it's formed in the vacuum than if it's formed in completely surrounded by water right because well the difference in the. Permittivity between vacuum and and water and so if a hydrogen bomb in the Purton is going to prevail. Then it better be quite dry. In water. Right. So although it may be structural determinant of a soluble protein it needs to be fairly dry. If it's going to become true structural that there a minute. So how does it. So you know collapse of the wind when the hydrophobic collapse takes place in the chain. Then you are also bearing the backbone. So not any kind of collapse qualifies as such because only hydrophobic collapse that good actually lead to the formation of backbone backbone hydrogen bonds is likely to prevail because otherwise the cost of the hydrating. Good be unbearably high compromising the stability of the compact structure. So the complex structure of a protein is achieved by delicate. Compromise between the number of hydrogen bonds that the backbone is able to make with itself and and the creation of a number of groups that eventually can actually separate themselves properly from water through this so-called hydrophobic effect. But so but then you may ask well how well may that all this hydrogen bonds. What are their chances that they will survive. Is it possible that activity the competitive at the needed to protect the bond. Could fail sometimes in some bonds. Creating certain structural vulnerabilities. For example if upon this part. Exposed the workers and then. Hydration make back on hydration may actually lead to a partial dismantling of the structure looking dismantling of a structure. So then well. So one way of assessing the micro biome end of these bonds is simply to count the amount of non-polar groups that surround them right so that would be much better than having to model their walker or whether water can get in or out. So you could just simply count the number of non-polar groups surrounding the bond within two spheres. Center they are for carbon So if the race. It is paired by the bond and then if you take another quick radius of those fears pretty much the cut of these things for four little statics in in water then then you can you can actually draw some statistics and determine how many numb polar groups from side chains are within those few years and then you have an idea of how dry it. Those hydrogen bonds are in a way how vulnerable to work terror attack. They are and them and then some quite striking features I rise from those statistics so for one thing the hydrogen bond is typically wrapped by. Nineteen plus or minus twenty six plus or minus seven dollar groups and then when you are in the tails of the distribution in the lower tail of a distribution you are dealing with them and the wrapped hydrogen bonds with a bond that can have enough. That's not try enough in water. It's a vulnerable hydrogen bomb and it turns out that those are sticky and they distinguish Parra logs so these are two essential features that make them quite an interesting target double feature for drag this guy. Very. Specially if you want to build. Specific drugs drugs with controlled and high specific city. So the idea would be the paradigm good be to try to the sign drugs that are actually contributing with non-polar groups to enhance their stability and their little statics of a pre-formed hydrogen bond this is the three body effect as you can imagine. So here is an example of this the hydrants as we call them hydrogen bombs that are prone to get further dehydration by dragging on nearby hydro Forbes in order to reduce their local dielectrics or in order to enhance the preform electrostatics So for example here in say here is a very curious feature of the P. fifty three. D.N.A. binding the main three as you know it's a crucial genic oncogene right because of its ability to police the the integrity of the D.N.A. and send the right or create the right a popped out the exact males in case the D.N.A. is not it's deficient Lee assembled and and so this is a region into digits with a major groove of the D.N.A. is actually a region deprived of tertiary structure. It has it's a helix that is sort of protruding from the structure like that but doesn't have a structure. So you can immediately sense that there's something wrong with this region right. In other words. If it doesn't have a tertiary structure that means it's probably not going to be able to wrap the the bands the hydrogen bonds that make the helix that are going to keep the killings in the place. So you know there's a quote unquote failure of copper. In this part of the structure or you may say there's a vulnerability there. And sure enough this backbone hydrogen bond is actually quite vulnerable to water attack but on the other hand. That worker is not really a happy camper. Because he has lost some core of the nation. Right. Instead of having called the nation for as symbolic It has now called the nation's three so that means this water molecule is probably quite easily expelled. It's there because there's room for it to stay there and hydrate the Carbon Hill but Iran i'm not be there right. So this is the hydrants are really. A region of tension. You could say of even interface shell tension in that the letter can get there but at a cost right at an interrupt because you know worse it's lost its core its full called the nation and at the same time the hydrogen bomb is not really happy there. So given the chance of a probe that this hydration process progresses then then the hydrogen bomb will be dismantled and they are my own company and we're just simply be fully hydrated. On the other hand if the right leader and comes seen then then that would actually MONTAGNE The structure because then you know you can actually if you exclude the water you can you can stop the lies the bond and also and hence it's electrostatic interaction so you know since we are looking at the Rugby League and as much as dielectric modulators here that that's sort of the paradigm we are after. And we and we believe that your pay. He that that this under wraps. The hydrogen bonds are actually creating the hydronic field which is the mechanical equivalent of the. Propensity to get to get dehydrated. Because as a hydrophone approaches it last words you would say in brought there. I mean in rather vague terms it lowers the dielectric of the permittivity and and that that would actually enhance the literal statics So this is not a conventional way of of of designing drugs because you see nobody would think of approaching the height of a top dollar but if your partner is interacting with another polar then the Herefore has a business approaching it because it can enhance the preexisting. Field Colombia interaction so is this the body effects are a little bit similar to what happens in human relations sometimes you don't like somebody but if a third person likes her then you start liking her too. Right. So it's a bit triangular. OK And so let me just now. Describe to you how these these vulnerabilities of a product in this they hide rooms behave in a pollution airy terms. So these they hide from patterns behave in a blue state. And so if you look at say a particular protein or function for example of a hydrophone the rate of days. Let's say a typical and Sime widely studied metabolic and some widely studied you look at it across species that have diverged billions of years ago and you notice something quite striking that pattern that always is always prevalent which is the larger. The population of the species the better May that the protein isso they fold the apology is pretty much the same right across species but they're wrapping is. Different the wrapping is getting worse. Right so sorry to say but our proteins regarded as subunits are allowed to see or than those of bacteria and which is in turn lousier than those of archaea. Why. Well essentially because we selection is far more inefficient in human than it is in bacteria or in archaea right we have a much smaller population and hands. Our probability of sixteen my little experience mutations in the population is much higher in human than he's in species with you know they're not of the magnitude larger population. Right. And so as a result our proteins are more loose. But that could be also a blessing in disguise because that means that this protein is far more reliant on binding partnerships to MONTAGNE structural integrity and in fact at first. There's those complex ations it promotes those complications because as I told you this back in the fix that the hydrants are actually sticky and hands because they promote their own the hydration The good thing to open up knishes for the development of protein protein interfaces right. So the fact that these protein is lousy and then this or more loose in a way than this one opens up the probability the possibility of higher level of complexity. So the origin of that complexity is actually none of that developed at all. It just them from the simple fact that selection is far more inefficient in in how you carry it's done. He's in prokaryotes every now here. So the same happens now. And so I examine what happens across all thought looks right at the same. They're in different species. Now what's of direct interest to the pharmacological industry is what happens across Perlozzo right. So high marks within the same species that's where the possible that's what we have to try to tell them apart because if Carson reactivity is her eyes that may lead to side effects which is precisely what we are trying to avoid. So in part of Los this story is a little different. So the parlor arose from a gene duplication event right. So a gene duplications event producer dosage imbalance because. Let's say the gene the product. Operates in that complex ation right. And now I have twice as much population or concentration of the gene product after the gene duplication event and that is not properly titrated stoichiometric least with the concentration of the binding partner which. That remains had the same concentration as before the gene duplication event took place so that creates had the city in balance. Now if the product in these highly rely reliant on binding partnerships to maintain the structural integrity. If it's not a loner. Well then that's going to have fitness consequences because now you have a perp thing that needs the partner but doesn't get it right doesn't get it in the right amounts and so and that's going to be quantified by the wrapping quality the poor the wrapping the more reliant it is on binding partnerships and the higher the fitness consequences. And so there are two solutions to that problem. So either. The one of the copies of the gene is simply selected out if selection is sufficiently efficient or so. Functionally zation will take place in other words there's going to be some level of power look divergence. Due to the accumulation of my little it eeriest mutations that's going to change the wrapping of one copy with respect to another or of the expression. But they're not of the past transcription a regulatory part thems that are recognized by the micro-mini But some level of they burgeoned has to occur to meet the gate the dosage imbalance. And we have we can harness on that divergence to create better drugs because we know that impart that by purchase is going to translate into differences in wrapping across panel looks right. I mean that's a fine tuning of different functionalities for Perth things of common ancestry and so and because we know that wrapping the fix are sticky and the Parlow have different patterns overlapping defects. We can create better locks right more specific dogs dogs that can actually tell apart different members of our family. Likewise because power loss will tend to express themselves in differently distinguish their expression pattern across the shoes to another where you're mitigating the dosage imbalance then that that opens up another opportunity for that act designer to create teacher specific delivery and so for example if you want to tell a part to power looks when you want to touch and the other you don't want to touch and the dog is not cannot operate to have the critic has to be cross reactive because well your drug happens to be cross reactive you can. Still managed to distinguish them by teacher specific delivery. Right. So you know that one part there when you want to touch express here but not there. And so you just direct your cursor reacted the rock to that particular tissue and then you ensure that you're not that you're going to be able to tell apart the pilots. So in essence the Parlak divergence arising. Essentially from the dosage imbalance and it's fist if it and its consequences can open up opportunities for better drug design. Based on these evolutionary observations So for example here is how it works. Remember I show you the two examples early on of the Focalin he's from kinase and the insulin receptor structure very similar to the room in square this placement a less than a man strong for the backbone for the backbone atoms but at different pattern of the hydrants at the active site so perhaps one could take advantage of this difference to create a bracket that can actually tell apart before collocation Tiny's uneven receptor and that would be of great interest to the pharmaceutical industry because it would enable you to treat. Tast asis without in the risk of inducing of A or B. to call my own the patient in the patient. So this is the part of the I'm although that's you have to excuse my pomposity here. Call it really paradigm. This is my suggestion. Let's call it like that for now. So they way people good go about expanding lead. Typically have relates to what kind of target of all groups there are on the surface of a protein that they can match. Right. So they say no there's a non-polar experiment. Non-polar group on the surface. So OK we'll try to put a non-polar on the on the drug right to match it and and to enhance affinity and that's fine. Nothing wrong with that except that these features are typically highly conserved across paradox. So if you try to target them or Likewise if there's a problem accept or you put a problem down or or problems are put upon accept or imply that that's conventional thinking what's wrong with that is that you are targeting features that are highly conserved and hence you're promoting promiscuity. So that's the so that's you see selectivity and I think that the are two completely different ball games. And you don't you don't want to target the long delayed hanging fruits of the forbidden fruit is here in our words because these are the obvious things that a pedestrian they sign are good do but they would lead to permit security and and so what I suggest to do is just try to introduce non-polar groups that would actually improve the wrapping of preexisting hydrogen bonds the hydrants which are not conserve across Parlow and Di could be a feature promoting specific cities. So this is how it works so. So for example let me just give you an example. So here are the crucial structural parts around the A.T.P. pocket over focal and he's been kind he's in the engine and receptor and you see in green you see an under wrap hydrogen bonnet the high don't and gray denotes I will wrap one. So there's the this is there's a wrapping difference here that distinguishes those two proud things. So we took an an inhibitor of the focal adhesion kinase produced by no part this call to eat two to six. So. This inhibitor is never going to see the light of day because it induces the Happy to combine the patient. So you're never going to see if the program but we just modify to doing in increasing the level of the wrapping capability for the for this particular the hydrant in the focal and he. Which as I told you I Lions with a well wrapped hydrogen bond in the engine receptor so that amounts to just do some synthetic chemistry starting with a different reactant and you end up having the mental what you wanted to be it to be. And here is the US A for the insulin receptor. Immune a chemistry for the insulin receptor which as you can see the theory compound. Knocks out first for a relation of the insulin receptor but our compound which just the first on the on the lawn the previous one in only one method doesn't touch things in receptor You see we don't touch it and this else. This article IO blastoma sells the kind of cancer that ended up killing Kennedy which are of course highly expressive of the insulin receptor right the brain uses a lot of a lot of glucose to function and and so all of this and Gaia blastoma uses glucose because he has metabolic from an overdrive it uses even more so. So he has a lot more insulin receptor and so these two type of low blastoma cell. You see here he knocks it off and knocks it out and our compound doesn't touch it. So and. If you look at the ovarian carcinoma cell line it's called which is another expression of fact this is a highly metastatic cancer. We. Inhibit force for relation to virtually the same extent as the parental compound. So we could say that we can tell apart. Perhaps we can tell apart Parra looks based on wrapping differences and this sign in drugs accordingly. So knowing going to describe in much more detail how this works very specific case so here is very the darling of drug. Of molecular intake cancer therapy in martini which is an inhibitor of two major kinases of folk the able some kind nice and the kid kinase and here. Here is either there are. Sitting in the wrapping part of the keep kinase and here are the backbone hydrogen bonds the drug is actually wrapping. OK It's actually contributing with non-polar groups to keep to stop the lies those plans. And so. The inspiration came from this paper which came in two thousand and six. That said that actually. Imagine it was kind of a toxic and traced its carrier toxicity to innovation of one of its major targets they pull some kind of this broad of course pandemonium because remember suddenly it turned out that all these edema that people were seeing in patients treated with Imagineer imagine there is a major drug to treat. Both just interesting I'll struggle to more and chronic myeloid leukemia. So this is the my was not due to the cancer but the drug itself and actually ended up creating a card there hypertrophy and all to mentally. A stroke and so. The idea worse. Can we really really sign imagine if to curb this side effect right. So if the wrapping technology or the wrapping paradigm is going to hold. It's going to be better tight then it better work in in we better take up the challenge. Ladies and sure that he really works. Or else you know we just just live in only in the theoretical space. So this is also interesting for for long term treatments where you know imagine it has been used to treat diabetes but they are of course the prolonging treatment makes only things worse. And so it's in our interest to try to curb the cardio toxicity you see in the wrapping really sign of the drug. So this leads us to the second problem when dealing with side effects which is the context dependence of the target so here is able some kind of which in the chronic myeloid leukemia cell progenitor cell is actually deregulated through some key marry. Interaction with the P.C. our genes and so this this kind is actually. Considered to be active all the time doesn't stop. And so it's inhibition really blocks entire pop thought that pathways these are well known popped out of pathways transduced originally by the able some kind. And that's exactly what you want. Right. You want to block any type of thought the primary was the one you want this cell to die. You want to in fact induce a pop doses. So this is so this is exactly what you want to though and that was the rationale behind imagining behind the discovery of the matinée But when you go to the heart. It's a different story. Here is the car you're myositis and they are the initial of able some kind. This actually triggers a cascade of forceful relational bends that ultimately leads to mitochondrial the polarization. And ultimately A.T.P. the policeman and you know that the card in my a psych cannot operate kind of therefore the A.T.P. the policeman because he needs A.T.P. more than any other cell in the body is working all the time right. So this this this type of stress opened up last Matic reticulum strace ultimately causes the end of the country mio side. So this is a side effect that we need to address. And in passing we notice that this cascade of events leads to the forceful relation of June. K. so what if we try to redesign the mat in a so that it doesn't touch the able some kind. But him passing we really rather also. Make it so that it inhibits Jim K. and then we'll be able to protect the heart because the pathway that is actually triggered by the stress. Good actually be blocked at this stage and then the mitochondria will be preserved. So essentially if you go to the drawing board. This is what you really want this is this Are the cross reactivity of imagine if this hard star gets and this is the therapeutic impact of the drug on the different targets and you see the key it is related to the is a major target in treating a solid tumor gastrointestinal stromal two more and perhaps even kid dependent melanomas although that's only in Phase two trial busier able kinase It's great to treat chronic myeloid leukemia as a great target but as strong recently it produces it induces cardiotoxic city and then period. Here far it's a plot to let the pending growth factor receptor kinase which of course its inhibition would be highly desirable because this is an input and eugenic factor so you want to cut the blood supply to the two or more. So it's OK that inhibited this course of activity in fact it's highly expected because these two are close homologues kit and so whenever you touch these you touch these L C K You want to really want to touch L C K. This is a limb for side dependent kinase and that. It's in the patient can actually produce immunosuppression suppression and you don't really want to touch that. So the desired targets would actually be keyed P.G. or far. And Jim K. right because we know there's a junk A is on the block in junk A can actually have a protective role on the mitochondria of country Meyer sites. So we compare the wrapping. Of the primary in the new targets busier able and Kate and we immediately notice that there's a difference here in the so-called nuclear type binding look so here you have a well wrapped hydrogen bond in enable sun which becomes the hydrogen keep it when you do the alignment. So that's a major rapping difference that we may want to take into account when redesigning the martini to be able to tell apart kit from Ableson right. Remember we want to avoid cardio toxicity. We want to create a new version of the mat and if that is not cut or talk sick the same is true about L C K. The same story. So keep it has of the hide on there is sick a has a well Rob hydrogen burn so what if I modify this term in a ring of a martini to make it a better wrapper all of this the hydrant. And then I in that way I could ensure that I'll be able to tell apart L C K From Keith and hence create a drag. That doesn't have the immunosuppression. So this is the way you really proceed. You actually have to digitalize this information and because you are dealing with para logs. This is the this is the the Hydra matrix as it were so what entry I'd say you have a color entry. If I enjoy a racy those are paired by the hydrant and great if they're prepared by a normal hydrogen bomb and nothing you have not paid out by a hydrogen bomb at all. Right. So you can actually compare the wrappings of these different targets by simply in this sort of digital form. And and so you can create a library of closely related power logs in order to be able to manipulate specificity. According to the road to the differences in the wrapping patterns across parallels. So here is of the hydra so well rob a hydrogen bomb enables on a well run hydrogen bomb in no sich a hands up possible candidate for how your specific city. Right. And so here is in the cartoon so you see these are two the hydrants A.M.B. in speech in protein one only A is pretty present in protein too but not in protein three so if you target it with a wrapping this sign and aimed at stopping lies in a way then chances are you're going to have cross or activity between one and two but not in three. If you target be on the other hand you're only going to get closer activity for one but not for two and three right. So that's the idea behind it. So that's just how you use your filter for selectivity in a digital manner. This adds. An extra twist to this story when we look at Jim K. junk A in the wild type doesn't even have the hide on there has nothing there. It's an open loop. Why this region is so under wraps. It doesn't even form a hydrogen bomb. Right. I mean if you have a list then seven. You don't you never find a hydrogen bomb with less than seven non-polar groups in the in the in the microenvironment around it. So if I mean right seven. It's another story but. It's so this is so poorly wrap this look that it's not even the hydrogen bomb is not even there. But when you bring in Greece. Here you can actually induce it. So if we if instead of using the Matheny you could use imagine it with a methyl here. Then you would actually induce the bond and and so that's there's a case of controlled induced folding that can actually be used to achieve specific city. So this story is very clear now we have to put a metal there. OK To wrap this guy which is unique to keep it and to junk a and to P.G. are far and in not where else to be found in one of the other targets so in that way we are going to actually unhinge this specific city in a way that is going to curb the potential side effects especially cardio toxicity of the parent compound right. So I went to Billboard man and sent home pain from M.D. Anderson Cancer Center and asked them to well took them to the mirror and drawled them with some money and so forth and told him that I wanted this compound to be made. So this is remember a modification of the money. Only the first from the parental company one methyl and I claim that this is not going to be cardiotoxic and is going to retain all the anti-cancer activity with parental compounds. Sounds like science fiction but are for you. Work. So this is called W B C Bill barman and saying humping attempt number four. And we can talk about the three earlier times that failed but that's another story and we're only. So so here is a company we made W.B.C. for again the first in a men still on a matinee. And this is a first this so here is the cell growth us a guest in this inner struggle to power cells which are highly key depended and you see an equivalent. Reaction and an equivalent in level up in the center of cell growth for both our compound and the commercial name is Gleevec So it's also known as living now here is the first surprise when you look at chronic myeloid leukemia cell. Which which is able some dependence line. This is the first difference. You don't see any in the vision of cell growth for when you use our compound as compared to what happens when you use clipping or the matinée you see. So this is the first hint that we may be achieving specificity. So then. This is a much more convincing test so here you have the cell the cellular assaye for the guts incision of terminal tumor cell with our compound wrapping read the sign and the Matheny and you see a comparable inhibitory activity over the phosphorylation kinase right. Now when you look at the chronic myeloid leukemia cell. There you see for a person. It's very different right. Our compound doesn't touch it. We don't touch Ableson right. We only deferring one missile but we don't touch a post. And Gleevec only matinees of course. Knocks down significantly the. Activity over the able some kind. Is it in Peirce it's phosphorylation significantly. Right. So we have we claim based on this our say that we may have achieved specificity. But is that enough to prove that we can really care. The side effect. OK So this is another I say so we gave it. We gave our compound and the prototype to a big corporation in San Diego California that can do extensive kinetic assets and this is what we got so here are a good portion of the humankind and there's another slide that shows the rest. So it's not the entire human kind but it's a good cross-section of it up at least two hundred fifty or sixty kinases out of the five hundred and twelve and nineteen that we have and so here is the able some money for able sun and some drug resistant mutations of able sun and you see our activity is. Sub micro Mauler where the micro Moller where the Matin of activity unable Sunny's typically and then animal a range which. And as you know this this activity is the culprit of the cardio toxicity of the matinée So we pretty much believe we have reduced it to significant levels because they are finity is only in the micro Mahler range which is not of four mycological significance. But what's more interesting though we don't touch L C K. Right here is they all stick a band for. Forty Matheny and there's not red here. We don't we don't really have any activity on L.C.. We have comparable activity in Kate and surprise surprise with touch junk A Remember this. The protective role of the heart. So we can actually get that. By the wrapping engineering of the martini and the world type. Excuse me the parental drug doesn't touch. Junk a right. And so. So we are to pretty much what we want it right. We touch keep it to the same extent us imagine it with hardly touch able some which helps us prevent cut it actually City and with touch junk A which had a protector role on the heart. By inducing the hydro where there isn't any and without that shell sick a there for our drug is not expected to be immunosuppressive. And as for P.G. as far. We have nothing to say it's not in the P.T.B. although we can construct a homeowner to model based on kit this close proximity with touch it to the exam extended to parental drug and that's OK because we wanted to in fact happen to Janick activity and we do. So now in regards to the to the side effects. So here you have the car the in my ear sight. Of So we need it. Of course color my our sites in we need to show that the Matin able Gleevec actually turn now on. They pathway that is turns do is through Jump A and eventually leads to mitochondrial the polarization and sure enough the bands are there. So this means that this pathway has been activated. Whereas our compound hardly touches it. In fact this shows inhibitory activity on junk a forceful relation. And then we have you see one methyl can really protect the heart so site of cytochrome C. is a typical mitochondrial protein. If it goes into the site of Solich space that means that that means big trouble. Right. That means that translate. Patient implies that the mitochondrial has been dysfunctional and sure enough look at this massive translocation of cytochrome C. to the site of Solich space and we hardly get any outward mitochondria is far more tight than that treated with a mat in it. It has damaged the mitochondria of the cut in my eyesight. So then the of course we needed the animal model. So we use this female not my eyes. That has no thymus And so to which the intestinal Stromile to more has been or thought topically induced in the central graph and. And so you see comparable activity in terms of entire cancer. In tight you more activity for both and so the untreated my eyes. You see the exponential growth all the way to more mass and so but then when you look at the curve the carrier toxicity The golden. Indicator the golden parameter here is the percentage of less than trickier ejection fraction and so is at ten percent decrease is of course lethal because it implies a massive this functionality in the heart and our compound even nothing else retains the same ejection fraction as the controls. If not improves it and that may be due to the protective role of ENTITLED TO MAKE A activity on the heart. So here is much more convincing picture perhaps. So here is the Mara over the over the heart of the bias in the treated with the matinees and B.C. for the Z. say in this scene a graph study four with a new my eyes see no graph just study gushing says he has terminal Q. more. Model. And so you see they live been trickle in the diastole and in the sisterly if nothing else the cross-section almost remains the same and is even compensated by a bloated Oracle. You see this heart is really not pumping well at all. In fact these mice had to be sacrificed actually died after the six weeks of treatment. With our compound that remember only the first one methyl from this one this this mice are are it's perfectly fine right. I mean this you see how tiny the cross-section of the Live been trickle it is in the sisterly right. So this is this is actually a functional pumping well pumping heart and and so I think I've tried to show you in this lights that you see in the wrapping concept. You can actually distinguish Parra looks in ways that will enable you to control specificity. So that you can curb side effects. By telling a part para los. So you may say yes well that's an example but this is universal so to what extent can we control specificity by using this wrapping concept right I mean the maybe that this was sort of this was a success story but you know just a product of luck rather than anything else. So so I decided I could just compare the wrapping patterns of the major drug targets with the with the pharmacological press file of those targets so you could actually compute the well like like to call the wrapping hull over of a protein. So the Robinho are all the the hydrophones that are actually wrapped. But you're driving so here is a drag. Here is the height on part of the drive and those are the B. hydrants are affected by that. Right right. That they drive where the drag is making a difference in terms of enhancing the local stability of those all those vulnerabilities. And so on then you can actually align those those hulls because these are again these are close parallel looks and look at the wrapping differences right across Parlow So how do you look at what you mean by wrapping distance well so for example. Remember the high don't part then can be encoded in a matrix right in a binary matrix one at position night Jay if i and j are P.R. by the hydro and zero otherwise. And then now. Now that each product A needs a matrix. You can actually take a coming these times of those matrices. And we call that the wrapping distance. I mean perfectly fine because if the structures are line level then the matrices are a line of zero and if you are not you can use. Well you can you have to think a little bit with them to compute those Hamming distance is but in principle you can do it. And this are all targets that have structure in the P.D.P. and these are the wrapping distance matrices matrix across those targets. For each for all those pairs right. Those pairs of protein targets and then you can independently look at the pharmacological this turns matrix so for each propane you have your affinity profile against a background of drugs that are successful. So here this we have to exclude star a sporting which is a highly promiscuous compound and it's not really an effective drug at all. And so it's for it. Then you can associate vector and then you can think the Euclidean distance. He affinity Victor's right. And so that that would give you an idea of the difference in pharmacology across to between two proc things right. So I mean the distance would be zero. If the two proteins react identically to a set of. Background set of drugs and and one in the opposite case in the other extreme and so you can define and actually have pharmacological distance matrix and then the question is OK how do the two of the Compare of the two of them compare and they actually correlate alarmingly well although none of those drugs were designed using the wrapping concept. I mean if they were designed using the wrapping concept then then then it would be completely tautological and the correlation would be absurd right. What I'm saying here is that these drugs that were found through trial and error and painstaking effort actually are essentially obeying the wrapping concept although they were never meant to be designed using the wrapping concept. And so now that we know what may make said drug successful we may use it rationally and intentionally to create better drugs right because as I said none of those drugs were were had the wrapping technology in mind but but this correlation bespeaks of the apparent universality of the concept in terms of trying to develop a new field there to achieve specificity based on a structure there I features of the purported targets. And with the. With that I would like to end. And thank my fine funding agencies and my group that it all the work and all these people that helped me think straight about these concepts and and also I want to thank you for your very kind attention. Yes. Crystal water yourself. Yeah so well that points to a big topic important problem what you're saying which is the you know normally these items are in the twilight zone between order and disorder or so what they're really they are induced friending features. So you know worse if you don't have a leak and chances are that water will get in and disrupt these bonds. So oftentimes what we see is if you have a poor wrapper or then and yet in a crystal structure you see water in third collating oftentimes the the idea is that. We don't really know exactly. I mean this is a pretty dynamic object you know worse when we say it's very under wrapped or it's partner able to water attack. What we are saying is a centrally we're introducing some underlying dynamics into the picture right. We are saying in the absence of a league and if you if you examine that particular panel look. Chances are that. That hydration of the backbone will prevail but given the right leg and that hydration is skin there and the structure is retained so. So we are in a way this is pointing to the problem of you know what constitutes the Falding the quality of a particular fault is it when we identify those interactions though those those packin defects are wrapping the face. If you wish we are actually that seems to be indicative of ephemeral adventitious or permanent complex ation So the structure has to be viewed in in the context of some level of complex ation in the absence of that level of complex H. and I don't think you really have a Luckily a structure there. It's. Again we can you can actually show that because you can infer the hydrants directly from sequence. By Because from sequence there's something you can infer with a good degree of reliability and that is the propensity for this or that there's this sort of a propensity right. Now with and now they're algorithms to to to infer the sort of propensity and we don't care about regions of certainty of these sort of there but we care about regions that are in between order and disorder and you can see that these sort of their profile across the sequence and what matters are the regions with high slope because those are those are regions of tamed. These are all the regions where it may be the sort of there but it's not a wild this or that or if you if you have. The right leg and you can actually maintain this structure. Alternatively regions are like there's very structure. For example a protruding piece of. Say the P fifty three D.N.A. binding the main are also very rich in the height and you know we'll see if there's no leg and not there. We'll just come in and hydrate the backbone and dismantle the structure or show off significance proud things that really cannot keep the structure at all. For example pro things so when you look when you ask yourself what is the most under wrap protein overall in the Pretty well it's a cellular structure there the the the cellular structure of a prion you know worse it hardly qualifies as a solvable protein is so vulnerable that qualifies as solar protein what most likely happens is it aggregates it goes into a different face it doesn't really cannot hold its mana Meric So actually not all other highly under wraps proteins are talk scenes for example they hold they hold this structure because they have a lot of die so five bridges. So it's a very dynamic concept where what the has a high propensity to get in and and this man. With. Was. Now. Well you know. This is a hole here. You guys rock you here stays exactly the same. But there's a whole structure. So I could show them that. Yes if you're right this is right. Then in the former case the feelings were not. Yes. And indeed indeed yes we have done those controls. Yes we have done them with with a Manton here. That's a good very good question and I think. There's a hole there there are several holes in the in here where there's this water. I believe it seemed this region so we have no it's not really so we have expanded. We have expanded the tail of the matinée in regions where there is where there is well there are scripts telegraphic water but not the hydro and there are whole in the structure as it were and we don't see any any difference in affinity. Yes we have than those controls and if you look different because science person asked if physical difference. X. Y. yes are other differences. Besides there are other differences in where there are differences. There's people have tried other ways of engineering specific city based on for example orientation of these highly conserved Phinney Lalan in for example. And this is the so-called This is activation look. And so there they have they have they have tried to look the differences in the confirmation of the Activation loop for example which are which distinguish kinases in the active state or in the not to excuse me now. Inactive they were in the active state. They all look alike but in the neck to stay there. There are differences and yes they they can they can engineer specifically the sound degree but not in a systematic manner. You know worse. It's not it's just a feature that may distinguish two particular kind eighty's. When there are many others but but not you can just have a universe of targets and say I want to work here here and I don't want to go there and there in a universal Manor. But there sure are differences in. I mean the region that is more prone to this sort of there is the region that is also more prone to variability. But it's also the most elusive region because it's more flexion and so chances are you're going to induce Falding there and that's I while problem as difficult as the protein folding problem. I mean to try to predict the fault or the induced fault right or where is it going to go. So. I cannot predict how we will escape inhibitor. I know and I can rationalize a post to reorient how it escaped inhibitor. But I cannot predict So for example. To this wrapping I don't know what we will do to this wrapping things probably will modify we haven't looked at that but we looked at how we escaped from the parental garage and sometimes this cave from the parental rights introduces mutations that produce new the hydrant and then we can take advantage of that wrapping difference in the target to chase. They are the nature. In that we can actually create a drug that has affinity for the moving target. For the drug reason for the drug for the matinée resistant Berry and now would be interesting to see how nature responds to the wrapping deciding you know was how does it escape. For example if you target we have we have tried that we have tried to target this the heightening the activation loop and and by putting a metal here and we are asking ourselves well what kind of some Matic mutations will ever arise here. Probably mutations that will wrap this properly and make it disappear. Or maybe something that even this stuff realizes they look further so that this doesn't form or All right so you don't even have the so it's so under wrap this region that now you don't even have a the hydrant in the in the wall in the in the there are a system mutation mutant but we don't know yet because they lack time for those experiments it's it's on a different scale. So we. It would be interesting to test what are the nature's strategies to escape wrapped in this science has. Yes. We do in particular we got this one. Now where is it just a minute for this is the most striking one. So we. We've got. You see junk A doesn't even firm up the hydrant. But if you bring enough green. To the surroundings. You can induce it and now we make a crystal structure for that and and they did I was to show that in those folding if it's controllable. In other words if it occurs in a restricted area I mean I don't talk about you know those falling an entire problem but if you can induce the heightening a look. Say you can really use that as a selectivity triggering feature. If you. Well if you do it properly understand the rules of the game. Right. So here what we're doing is we're just trying to wrap it a little bit there so that we can actually form and that had that was initially shown in simulation and then crystal structure revealed that the loop closes up. But we don't have for example the crystal structure of the of a matinee of our rapping version of the matinee with the targets. Unfortunately it just takes forever. Sometimes doesn't work or. Yes. We use exactly the same constant we use exactly the same concentration in this experiment than we use for the matinée that we use pretty much exactly the same which is. Five milligrams per kilogram or something like that. And so this this process are made exactly the same concentration and so you see the tumors taking off in the central graft and and this is what's so this experiment. So for example if you. You use the same concentration as Imagine if the mice died in six weeks. This mice live for about thirteen or fourteen weeks. And you know which seems to be the life cycle for Newt mice. So in fact they were they were OK. As far as we can see. Yes or. Yes Well very important. Well for example if you look at our proc thing the hurdles are particularly prevalent in nucleic acid protein interfaces. And the reason for that seems to be C. This is a protein. This is a region of a protein which interacts with the D.N.A. Right and. And this is full of the heightens full of the hydrogen and if you look at transcription factors. They're full of the hydrogen The reason seems to be that they promote the hydration and that's exactly what you need to get the right a little to get a little statics between the D.N.A. and the Argentines of the protein go in. You know which not only you need the little started marching. But you need a reason to exclude the water from the interface. To get it going. So Sol this. This sort of micro cavity what it can get there and actually you see if they are genius were planted in affluent surface. Then they would not be as effective or license say because they will be hydrated. But if they are dick or a thing a region where water wants to get out then that's much better. As far as enhancing the literal statics of the recognition of the phosphates in the major group. Or when the nuclear gives it itself. Well that's something we haven't explored. But maybe it points to thing to thing bass tacking east use Right I mean why do you have higher stability when you have things that are stacked well. You can argue many ways in this there's been a whole. I mean there's a whole theory of that semen and all these people but one argument is that they protect each other right they exclude water from one another and so stacking actually enhances the illiterate statics but we haven't really explored the hydrogen nuclear but certainly we have examine D.N.A.. Protein interfaces and those are extremely rich in the hydrants. Thank you thank you.